Nu, nu&#39;-bis (alpha-sulfoalkyl) actinospectacins and process for their production



United States Patent 3,147,277 Na -Bi8(a-SULFQALKYMACTINQSPECTACINS AND PRQCESS FCR THEIR PRODUCTION Herman Hoelrsema, Kalamazoo, lllfiich, assignor to The Upjohn Company, Kalamazoo, Mich, a corporation of Delaware No Drawing. Filed Feb. 9, 1962,8er. No. 172,099

8 Claims. (Cl. 26i)340.3)

This invention is related to novel compositions of matter and to a process for the preparation thereof, and is particularly directed to novel N,N'-bis(a-sulfoalkyl)aotinospectacins and to a process for producing the same.

Actinospectacin is a biosynthetic product produced by the controlled fermentation of Streptomyces spectabilis. Various methods for the production, recovery, and purification of actinospectacin are described in the published literature, e.g., D. J. Mason et al., Antibiotics and Chemotherapy, 11, p. 118, 1961; M. E. Bergy et al., Antibiotics and Chemotherapy, 11, p. 661, 1961, Union of South Africa Patent No. 60/ 4,098, and Belgian Patent No. 596,175. Actinospectacin is characterized by an optical rotation [a] =20 (H O); by solubility in water, methanol, and ethanol, and by insolubility in acetone and hydrocarbon solvents; and by the presence of two basic groups: pKa 6.95 (E 0), pKa 8.70 (E 0). It was formerly thought that the molecular formula was wherein R is selected from the group consisting of hydrogen and lower alkyl, e.g., methyl, ethyl, propyl, butyl, and the isomeric forms thereof and M is a cation, for example, hydrogen, alkali metal, alkaline earth metal, ammonia, magnesium, and the like.

The novel compounds of the invention, N,N-bis(lower a-sulfoalkyl)actinospectacins are obtained when actinospeotacin is reacted with an alkali metal bisulfite and a lower alkanal, advantageously in the presence of a strongly basic anion exchange resin. (Suitable anion exchange resins for this purpose are obtained by chloromethylating by the procedure given on pages 88 and 97 of Kunin, Ion Exchange Resins, 2nd ed. (1958), John Wiley and Sons, Inc., polystyrene crosslinked, if desired, with di- 3,147,277 Patented Sept. 1, 1964 vinylbenzene prepared by the procedure given on page 84 of Kunin, supra, and quaternizing with trimethylamine, or dimethylalkanolamine by the procedure given on page 97 of Kunin, supra. Anion exchange resins of this type are marketed under the trade names Dowex 2, Dowex 20, Amberlite IRA-400, Duolite A-102, and Permutit 8-1.) For example, when sodium bisulfite and formaldehyde are reacted with a mixture of actinospectacin and Amberlite IRA-400 resin the novel compound N,N-bis(sodium sulfomethyl) actinospectacin is obtained.

The novel compounds of the invention, N,N'-bis(oz sulfoalkyl)actinospectacin have activity against bacteria. N,N'-bis(sodium sulfomethyl)aotinospectacin, for example, has the following antibacterial spectrum when tested on agar plates by applying 0.08 ml. of a 2 mg./ml. solution of N,N'-bis(sodium sulfomethyl) actinospectacin on a /2" disc.

Microorganism: Zone size, mm. Bacillus subtilis 25 Staphylococcus aureus 24 Klebsiella pneumoniae 26 Sarcina lutea 17 Proteus vulgarzs 22. Salmonella gallinarum 19 Proteus rettgeri 22 Thus the novel compounds of the invention, N,N-bis- (lower a-sulfoalkyl)actinospectacin are useful to control the infection of silk worms caused by pathogeniccultures of B. subtilis; they can also be used to minimize or prevent odor in fish and fish crates caused by this organism. They are also useful as oil preservers, for example, as bacteriostatic agents to inhibit the growth of cultures of Proteus vulgaris that contaminate used cutting oils; they can also be used to prevent or minimize the liquefaction in packed yeast caused by this organism.

The following examples are illustrative of the process and products of the present invention, but are not to be construed as limiting.

Example 1 .-N,N-Bis(S0dium Sulfomethyl)Actin0- speotacin A 3.5 g. quantity of actinospectacin free base in 35 ml. of water was stirred at 4 C. 1 /2 hr. with 10 g. of a strongly basic anion exchange resin. (The anion exchange resin used for this purpose was obtained by chloromethylating by the procedure given on pages 88 and 97 of Kunin, Ion Exchange Resins, 2nd ed. (1958), John Wiley and Sons, Inc., polystyrene crosslinked with 5 percent of divinylbenzene prepared by the procedure given on page 84 of Kunin, supra, and quaternized with trimethylamine by the procedure given on page 97 of Kunin, supra.) To this mixture was added 1.5 m1. of 36% formaldehyde and 1.88 g. of sodium bisulfite. After 16 hrs. of storage at room temperature, the solution was stirred with carbon, filtered, and then freeze dried to yield 5.06 g. of N,N'-bis(s0dium sulfomethyl)aotinospectacin having a melting point of 245 C. and the following elemental analysis:

Calculated for C H N O S Na -3H O: C, 31.1; H, 5.20; N, 4.52. Found: C, 30.41; H, 5.27; N, 4.55.

Example 2 By substituting the formaldehyde in Example 1 by acetaldehyde, propionaldehyde, butyraldehyde, and valeraldehyde there is obtained the corresponding N,N-bis- (sodium a-sulfoethyl)actinospectacin, N,N-bis(sodium ocsulfopropyl)actinospectacin, N,N'-bis(sodium a-sulfobuty1)actinospectacin, N,N-bis (sodium u-sulfoamyl) actinospectacin.

I claim:

1. N,N-bis(a-sulfo lower alkyl) actinospectacin of formula I CHaN CH3 S i HO 0 wherein R is alkyl of not more than 4 carbon atoms and M is hydrogen.

2. N,N'-bis(a-alkali metal sulfo lower aIkyDactinospectacin of Formula I wherein R is alkyl of not more than 4 carbon atoms and M is alkali metal.

3. N,N'-bis(a-sodium sulfo lower alkyl)actinospecta- 4 cin of Formula I wherein R is alkyl of not more than 4 carbon atoms and M is sodium.

4. N,N'-bis(su1fomethyl)actinospectacin of Formula I wherein R is methyl and M is hydrogen.

5. N,N-bis(alkali metal sulfomethyl)actinospectacin of Formula I wherein R is methyl and M is alkali metal.

6. N,N-bis(sodium sulfomethyl)actinospectacin of Formula I wherein R is methyl and M is sodium.

7. A process which comprises reacting actinospectacin with an alkali metal bisulfite in the presence of a lower alkanal of not more than 5 carbon atoms, and a strongly basic anion exchange resin to produce N,N-bis(a-a1kali metal sulfo lower alkyl) actinospectacin.

8. A process which comprises reacting actinospectacin with sodium bisulfite in the presence of formaldehyde and a strongly basic anion exchange resin to produce N,N'- bis (sodium sulfomethyl) actinospectacin.

References Cited in the file of this patent Patnaik et al.: J. Indian Chemical Society, vol. 30, pages 577-582 (1953).

Mahapatra et al.: J. Indian Chemical Society, vol. 30, pages 563-567 (1953).

Bergy et al.: Antibiotics and Chemotherapy, vol. 11, pages 661-664 (1961). 

1. N,N''-BIS(A-SULFO LOWER ALKYL)ACTINOSPECTACIN OF FORMULA 